Marcos Conceptual y Categorial

Australia is one of the world’s largest livestock producers, with the red meat livestock sector worth A$9.6 billion (ABARE, 2009) being a major component of the economy in both domestic and international trades. The average Australian consumes 46.5 kg of red meat per annum and 10.8 kg of this is sheep meat with 2 kg as mutton (MLA, 2014). This consumption equates to approximately 1.4 serves of lamb meat per week making lamb a significant component of the Australian diet.

Page | 16 The industry value and significant domestic consumption of lamb has led to a need to combat the possible negative health associations of red meat by some

consumers. It has been suggested that a diet rich in red meats may increase the risk of cardiovascular disease and colon cancer, which has in turn led to a negative perception of the role of red meat in health and well-being (McAfee et al., 2010a). In the modern western diet, red meat and animal derived foods are a major component of human nutrition. Similarly, in developing countries where affluence is increasing, a trend of increased red meat consumption is occurring (Myers and Kent, 2003). As a result of this increased intake, many nations are concerned for future population health burdens associated with high red meat diets. This could ultimately place pressure on already strained health systems. Therefore increasing the human nutritional health benefits of red meat through genetics and feeding may decrease the risk of chronic disease in diets which are high in red meats (Givens et al., 2006).

Saturated fatty acids (SFA) are considered one of the main health concerns and SFA is relatively high in red meat due to biohydrogenation of diet derived polyunsaturated fatty acids (PUFA) which is exacerbated by acidification of the rumen in grain fed animals (Bauman et al., 2003; Noble, 1981). Consumer education and retail training for “lean lamb” was used to reduce the amount of adipose fat on the average cut of Australian lamb which commenced in the early 1990’s. It is now reported consumers in Australia are eating heavily trimmed, leaner cuts of red meat. This trend no longer makes lamb a major dietary source of SFA (Williams and Droulez, 2010). As a result, consumers are now eating higher levels of intramuscular fat (IMF) which contain different fatty acid profiles and nutrients compared to older high adipose fat containing cuts (Williams and Droulez, 2010).

Page | 17 The omega-3 long-chain (≥C20) polyunsaturated fatty acids (LC omega-3) are

essential for human health and wellbeing. The two main LC omega-3 are largely marine-derived and are eicosapentaenoic acid (EPA, 20:53) and docosahexaenoic acid (DHA, 22:63). These two LC omega-3 have many scientifically proven health benefits for humans (Howe et al., 2006). EPA and DHA are not produced by the human body and need to be ingested. Humans do, however, have the ability to elongate the shorter chain (≤C18) omega-3 alpha linolenic acid (ALA, 18:33) to the LC omega-3 - EPA and DHA - but at an efficiency of approximately 5% in males and 5.5% in females (Burdge, 2004). Therefore, direct intake from dietary sources is considered a more efficient means at achieving the recommended daily intake levels of EPA + DHA.

It has been reported by Howe et al. (2006) that the majority of Australians are not consuming the recommended daily intakes of LC omega-3 and that dietary intakes therefore require significant improvement. The majority of LC omega-3 currently consumed by humans is sourced from marine based resources which are under increasing pressure from overfishing, climate change and competing industries (Nichols et al., 2010). Hence, new alternative dietary sources for consumers are of great interest to both industry and consumers (Nichols et al., 2010). Sheep have the ability to convert ALA to LC omega-3 at a similar efficiency to humans (Mortimer et al., 2010) and will deposit LC omega-3 as intramuscular fat therefore providing an alternative, non-marine source (presently at generally low content compared to seafood and other marine sources) of dietary LC omega-3. Pasture reared sheep meat contains dietary “source” (defined as 30 mg/135 g) content of EPA + DHA. These two fatty acids have abundant evidence for positive health benefits for

Page | 18 fatty acid ALA, however, with increased climate variability, it is expected that the availability of green grass will become more limited for prolonged periods and drought-affected sheep have shown significantly lower content of LC omega-3 compared to pasture reared animals.

Previous studies have demonstrated that the intramuscular fat content of LC omega- 3 in lamb meat is highly diverse and is affected by a number of factors with diet being identified as a key variable (Wood et al., 2008). The majority of lamb in Australia is reared on pasture and specialist fodder crops which results in lamb containing low but extremely varied, content of intramuscular EPA and DHA (Mortimer et al., 2010; Pannier et al., 2010; Warner et al., 2010). It is evident from prior Australian studies that the availability of green grass is a major factor in

determining LC omega-3 content in Australian lamb due to the abundance of the LC omega-3 precursor ALA (Bignell et al., 2011; Mortimer et al., 2010). It has recently been reported that lamb can meet the FSANZ claimable “source” content of 30 mg / 100 g EPA + DHA based on a 135 g serving of lamb entirely reared on irrigated grass at Cowra NSW (Mortimer et al., 2010).

It is well documented that the supplementation of lamb with rumen protected omega- 3 or feeding fish oils rich in EPA + DHA will increase IMF EPA + DHA content, but such an approach has not been adopted by industry (Kitessa et al., 2001; Wachira et al., 2002; Noci et al., 2011). Hence finding an alternative method to enhance EPA + DHA content which is readily adoptable by industry is needed and use of an ALA supplement such as canola, linseed or lupin may offer one approach. The cheapest and most abundant source of ALA for grazing livestock is green grass. However, climatic and production limitations results in the majority of Australian lamb typically grazing dry grass from late summer through to early autumn unless augmented with

Page | 19 irrigation and specialised fodders (Duddy et al., 2005). Alongside these typical

conditions, the Australian climate is changing and it is predicted extreme seasonal variation will become more frequent with expected increasing periods of drought, prolonged dry periods and higher temperatures occurring in many livestock production regions (PMSEIC, 2007). As a result, the availability of green grass to grazing livestock will be diminished significantly or quality reduced for increased periods of time due to moisture and heat stresses. Therefore drought-affected animals will have to be supplemented more often to meet their daily energy requirements. Given the importance of consumer perception of lamb meat as a healthy and alternative dietary source of LC omega-3, it is pertinent to understand options of supplementation available to maintain the LC omega-3 content of lamb meat during drought periods.

This study is the first attempt at providing a detailed assessment of the impact of drought on LC omega-3 content of Australian lamb raised under drought stress conditions, including with careful examination of the possible influences of breed, sex, genotype, supplementation and environment.

Chapter 2 examines the hypothesis that supplementing animals from a low intake of ALA (drought stressed irrigated pasture) with LA and ALA containing canola meal and lupin meal would improve EPA + DHA content in Australian lambs to dietary “source” levels. Understanding the effects on long-chain omega-3 content of finishing lambs with LA and ALA containing supplements - canola meal and lupin - in a 9 week feeding trial was the aim of this experiment.

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Materials and Methods

Animals and experimental design

All animals and procedures utilised in this study had the University of Tasmania Animal Ethics approval (A0009811) and were conducted in accordance with the 1993 Tasmanian Animal Welfare Act and the 2004 Australian Code of Practice for the Care and Use of Animals for Scientific Purposes. A half-sib experimental design was utilised. Five top-EBV rams acquired from Tasmanian Sheep Stud Breeders comprising Dorset, Texel, White Suffolk, East Friesian and Coopworth were mated to purebred Merino ewes at a ratio of 1:120 ewes in separate paddocks in a

commercial farming operation in the Coal River Valley, Tasmania, to generate 500 first cross prime lambs.

Animal management

Lambs were marked, vaccinated, electronically tagged at 6 weeks of age and run as one mob per sire group in separate paddocks within the same large scale

commercial farming operation under similar management conditions to minimise environmental variation. The flock was raised during a difficult season of severe drought with restricted irrigation capacity. From the third trimester of pregnancy onwards, the animals were raised on a mixture of limited irrigation, drought-affected rye grass pastures without clover and were supplemented with barley. At seven months of age, a representative sub sample of 40 animals with a mean liveweight of 32 ± 2.2 kg and body condition score of 3 were relocated for a 48 day feeding trial.

The animals were individually kept in 0.6 m x 1.2 m metabolic crates in an animal house at Cambridge, Southern Tasmania. Forty sheep comprising of 8 sheep from

Page | 21 each of the 5 sire breeds and assigned to two supplementary feeds (canola or lupin) and two feed levels (1% or 2% of body weight) in which ewes and wethers were equally represented within each sire breed and treatment group. Two animals were removed during the trial for health issues which resulted in a total of 38 samples being collected.

Ration composition

Feed rations were formulated to provide an isocaloric and isonitrogenous basal ration . The rations consisted of 500 g raw barley, 100 g chopped barley straw with a molasses spray at baling, 10 g of multivitamin mineral mix and either 500 g (1%) or 1000 g (2%) of each supplement of canola meal or cracked lupin. All animals had ad

libitum access to drinking water. Residual feed were recorded and discarded each

day and fresh rations mixed for each treatment. Nutrient composition of the experimental and basal rations are depicted on Table 2.1.

Blood sampling

Blood sampling was by jugular venipuncture directly into vacutainers containing EDTA, centrifuged and serum separated from the plasma.

Slaughter and meat sample collection

The prime lambs were slaughtered at Tasmanian Quality Meats at Cressy as per commercial Australian abattoir standards and using the same protocol as JBS Swift kill staff. Carcasses were chilled overnight and freighted to Wurhsthaus Butchery in Cambridge, Southern Tasmanian for full carcass breakdown. Longissimus dorsi

muscle tissue samples from 38 prime lambs were collected and transported to the laboratory in ice-containing baths and stored at -20°C until ready for genomic DNA and lipid extraction.

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Liveweight data

Liveweights were measured monthly using a Ruddweigh 3000XT walk over weighing electronic scale with capability of automatic scanning of lamb identity and

downloading of weight data into excel spreadsheets. Daily liveweight gains were then calculated based on final live weight minus initial live weight and divided by number of days in the trial (n=48).

Fatty acid analysis

Longissimus dorsi muscle samples (approximately 1 g) from the 12th _{rib interface}

were used for fatty acid analysis. Lipid was extracted using a modified Bligh and Dyer protocol (Bligh and Dyer, 1959). This involved a single phase extraction,

CHCl3/MeOH/H2O (1:2:0.8, by vol.), followed by phase separation to yield a total lipid extract (TLE).

An aliquot of the TLE was trans-methylated in methanol: chloroform; hydrochloric acid (10:1:1, v/v/v) for 2 hours at 80°C. After addition of water, the mixture was extracted three times with hexane:dichloromethane (4:1, v/v, 3x) to obtain fatty acid methyl esters (FAME) which were concentrated under a stream of nitrogen gas.

Samples were made up to a known volume with an internal injection standard (19:0 FAME) added and analysed by gas chromatography (GC) using an Agilent

Technologies 7890A GC (Palo Alto, California, USA) equipped with an Supelco Equity-1 fused silica capillary column (15 m×0.1 mm). Helium was used as the carrier gas. Samples were injected, by using a split/splitless injector and an Agilent Technologies 7683B Series auto-sampler operated in splitless mode, at an oven

Page | 23 temperature of 120 °C. After 1 minute, the oven temperature was raised to 270 °C at 10 °C per minute and finally to 300 °C at 5 °C minute which was held for 5 min.

Peaks were quantified by Agilent Technologies GC ChemStation software (Palo Alto, CA, USA). Individual component identification was confirmed by mass spectral data and by comparing retention time data with those obtained for authentic and

laboratory standards. GC–mass spectrometric analyses were performed on a Finnigan Thermoquest GCQ GC–mass spectrometer fitted with an on-column injector and using Thermoquest Xcalibur software (Austin, TX, USA). The GC was fitted with a capillary column of similar polarity to that described above. GC peak areas were converted to mg/100 g using the 19:0 FAME internal injection standard prior to statistical analysis.

Statistical analyses

Fatty acid data were analysed for the fixed effects of sex, supplement, level of supplementation, sire breed, SNP genotype and their second order interactions using both generalised (PROC GLM) and mixed (PROC MIXED) linear model

procedures (SAS 2009), while the partial regressions of sire and herd were fitted as random effects. Least square means of fixed effects were obtained and tested for significance using the Tukey-Kramer adjustment test of paired values.

The full model was

ijklm kl jl jk il ik ij l k j i ijklm e H H b S S b SBSNP SGSNP SGSB GSNP GSB GSG SNP SB SG G Y                2 2 2 ) ( ) ( ) ( ) ( ) ( ) ( ) ( ) ( 1

where Yijklm is the ijklmth observation of the dependent fatty acid with fixed effects of

Page | 24 (k=1,2,3,4,5), SNPl of lth _{SNP genotype (l=1,2), first order interaction effects (GSG)ij,} (GSB)ik, (GSNP)il, (SGSB)jk, (SGSNP)jl and (SBSNP)kl of gender and ration size, gender and sire breed, gender and SNP genotype, ration size and sire breed, ration size and SNP genotype and sire breed and SNP genotype respectively. b1 and b2 are partial regression coefficients of sire and herd respectively, 2

1(S S)

b  and 2

1(H H)

b  fitted as random effects, and eijklm is a residual error term normally and independently distributed. All non-significant interactions were later removed from the final model.

Results

The 2% canola (2C) treatment demonstrated the highest average daily gains (ADG) of 112.5 g/day and total liveweight gain of 5.4 kg for the duration of the experiment. In contrast the 2% lupin (2L) treatment had a very low ADG of only 10.1 g/day and animals only gained 1.6 kg (Table 2.2). The 1% treatment groups were similar in growth performance characteristics (Table 2.2).

Fatty acid profiles of the Longissimus dorsi muscle tissue are summarised in Tables 2.4 and 2.5 (data expressed as mg/100 g). The overall trends of fatty acid content is broken down into the three major fatty acid groups of saturated (SFA),

monounsaturated (MUFA) and polyunsaturated fatty acids (PUFA). Sex, breed, level of supplementation and supplement type did not significantly affect the total

intramuscular fat (IMF). There was only a slight variation of 0.3% in mean IMF between supplements. Lupin supplemented White Suffolk had the lowest IMF of 2.7±0.6% and lupin supplemented Coopworth and canola supplemented East Friesian had the highest IMF of 4.3 ± 0.6% and 4.3 ± 0.8% respectively.

Page | 25 SFA was the major saturated fatty acid group present with MUFA content also very high (Table 2.6). Palmitic acid (16:0) was the major saturated fatty acid present in all animals. Overall lupin supplemented animals contained 16 mg/100 g more palmitic acid than canola supplemented animals, however, total SFA was almost identical across supplement types (Table 2.6). The minor fatty acid arachidic acid (20:0) was highly significant (P=0.02) for breed with Texel having very high levels present in the canola supplement group. Sex also had a significant (P<0.04) effect on arachidic acid content, with lupin fed males having very low concentrations.

Total MUFA was almost as abundant as SFA (Table 2.6) for both treatments, with a mean MUFA:SFA ratio of 0.97 for the canola and 0.79 for the lupin treatments. However, across breeds there was some variation. Coopworth sired lambs fed canola and their White Suffolk counterparts fed lupins contained more MUFA than SFA overall. The high MUFA:SFA ratio in Coopworth when supplemented with canola (1.06) fell to 0.63 when supplemented with lupin.

MUFA content also increases with supplementation in contrast to drought animals, but like SFA is not significantly affected (P>0.05) by supplement, sex, breed or ration size. Although not statistically significant there is a 163 mg/100g IMF MUFA content difference between canola and lupin treatments (Table 2.7), which may have

become significant if the experiment continued. Within the individual MUFA, only one was statistically significant, 18:15c (P=0.02). 18:15c was present at very low content in the IMF and males fed lupin had a lower IMF concentration of this FA.

PUFA content increased with supplementation in grazing animals compared to drought affected animals in the same flock. Total PUFA was very nearly significantly affected (P<0.05) by sex (P=0.052) and the IMF contents of a number of individual LC omega-3 were significantly affected by sex and breed. Total omega-6 PUFA was

Page | 26 significantly affected by breed (P<0.02) with Coopworth having the highest levels (150 mg /100g) than all other breeds and Texel having the lowest (121 mg /100g). The omega-6 PUFA - 18:26 (P=0.04) and 20:36 (P=0.03) - were significantly affected by breed with Texel having significantly lower contents of these two FA compared to other breeds. Sex was significant across a number of PUFA with females demonstrating greater PUFA content and in particular total omega-3 (P<0.05). The fatty acids 20:53, 20:36, 22:63 and 22:53 were all higher in females across the experiment. Ration size or supplement type did not demonstrate a significant (P<0.05) effect on any of the fatty acids. A further PUFA, 20:26, was also present at a very low concentration with a maximum of 1.1 mg/100 g and was statistically significant (P=0.04).

Table 2.1 Dietary composition of feed formulation components.

Nutritional Component Canola Meal Cracked Lupin Barley Barley straw + molasses Dry Matter (%) 96.3 93.3 92.0 92.5 Crude Fibre (%) 13.8 15.7 4.6 41.3

Neutral Detergent Fibre (%) 18.9 25.0 14.4 66.4

Acid Detergent Fibre (%) 15.9 20.9 5.5 43.4

Metabolisable Energy (MJ/Kg) 14.9 12.2 13.2 7.3 Digestible Energy (MJ/Kg) 277.3 183.7 213.3 62.3 Feed Digestibility (%) 60.0 40.0 60.0 20.0 Nitrogen (%) 5.3 4.8 1.7 1.0 Crude Protein (%) 33.3 30.1 10.4 6.2 Fat (%) 15.8 6.0 2.3 1.0 Ash (%) 5.9 2.7 2.5 9.6

Shorter chain omega-3 precursor fatty acid ALA

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Table 2.2 Mean performance characteristics of lambs by treatment group n=38 (mean values ± standard errors).

Treatments

Variable 1% Canola 2% Canola 1% Lupin 2% Lupin

Initial Liveweight 35.1 ± 0.4 35.4 ± 0.3 33.1 ± 0.4 34.3 ± 0.5 Final Liveweight 39.2 ± 0.5 40.8 ± 0.5 37.8 ± 0.6 35.9 ± 0.2 Total Feed Intake (Kg) 39.7 ± 0.8 45.1 ± 0.7 38.5 ± 0.7 39.4 ± 0.6 Daily liveweight gain (g /day) 85.4 ± 5.0 112.5 ± 5.1 97.9 ± 3.3 33.3 ± 10.1

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Table 2.3 Statistical significance (p-values) for the effects of sire breed, sex, supplement and level of supplementation on muscle fatty acid profiles

Fatty Acid Breed Sex

Supplementation

level Supplement Type

14:0 0.08 0.21 0.72 0.52 15:0 0.55 0.81 0.72 0.93 16:17c 0.03** 0.06 0.89 0.17 16:17t 0.44 0.09 0.28 0.26 16:0 0.20 0.14 0.72 0.38 17:0 0.10 0.05 0.86 0.54 18:36 GLA 0.71 0.43 0.95 0.76 18:43 0.30 0.07 0.27 0.85 18:2 0.30 0.35 0.12 0.70 18:26 LA 0.04* 0.16 0.45 0.47 18:33 ALA 0.83 0.31 0.65 0.68 18:19c OA 0.37 0.55 0.73 0.26 18:17c 0.07 0.09 0.09 0.97 18:17t 0.20 0.38 0.19 0.89 18:15c 0.12 0.04* 0.44 0.46 18:0 0.35 0.34 0.92 0.64 20:46 ARA 0.18 0.17 0.83 0.13 20:53 EPA 0.40 0.04* 0.83 0.79 20:36 0.03* 0.03* 0.94 0.97 20:43 0.31 0.88 0.25 0.90 20:2 0.64 0.02** 0.33 0.70 20:19# 0.12 0.08 0.09 0.71 20:17c 0.17 0.20 0.56 0.62 20:0 0.02** 0.04* 0.30 0.95 22:63 DHA 0.77 0.05* 0.82 0.49 22:46 0.53 0.43 0.66 0.57 22:53 DPA 0.56 0.01*** 0.93 0.82 22:0 0.30 0.54 0.74 0.89 23:0 0.88 0.80 0.49 0.81 24:19c 0.87 0.58 0.67 0.30 24:0 0.37 0.73 0.16 0.86 SFA 0.24 0.19 0.86 0.89 MUFA 0.44 0.41 0.84 0.97 PUFA 0.11 0.05 0.60 0.51 EPA + DHA 0.64 0.02** 0.87 0.34

EPA + DPA + DHA 0.66 0.02* 0.94 0.57

Total 3 0.72 0.05* 0.78 0.92

Total 6 0.02** 0.09 0.58 0.90

6: 3 Ratio 0.48 0.20 0.96 0.88

% IMF 0.19 0.43 0.27 0.52

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Table 2.4 Mean intramuscular fatty acid content (± standard errors) for each sire group and supplement type in mg /100g of the longissimus dorsi.

Fatty aci d 14:0 32.8 ± 11.3 56.7 ± 11.1 34.1 ± 10.3 29.7 ± 5.0 44.4 ± 10.1 36.3 ± 7.3 47.4 ± 2.5 32.2 ± 7.7 35.6 ± 4.9 24.0 ± 6.8 15:0 5.1 ± 1.7 9.4 ± 4.5 8.9 ± 4.0 4.4 ± 1.2 7.4 ± 1.7 2.7 ± 1.0 6.4 ± 0.9 7.4 ± 3.6 7.6 ± 1.5 5.9 ± 3.1 16:19c 4.8 ± 1.5 7.3 ± 1.0 6.3 ± 1.0 4.2 ± 1.2 6.0 ± 1.3 3.6 ± 1.2 4.9 ± 0.5 3.7 ± 1.4 4.9 ± 1.2 4.0 ± 1.0 16:17 25.4 ± 8.6 36.4 ± 6.0 32.1 ± 5.3 27.4 ± 6.7 29.3 ± 5.0 30.3 ± 3.8 23.8 ± 1.8 22.1 ± 3.7 18.7 ± 2.1 20.0 ± 5.0 16:0 381.6 ± 84.7 572.6 ± 91.0 484.5 ± 60.3 416.2 ± 102.7 419.2 ± 72.6 398.8 ± 48.0 404.3 ± 24.8 346.8 ± 50.2 331.0 ± 51.2 372.9 ± 84.9 17:0 46.1 ± 7.2 62.2 ± 4.3 55.4 ± 6.7 48.3 ± 10.4 40.2 ± 7.9 40.4 ± 7.8 47.4 ± 4.8 43.2 ± 9.3 37.9 ± 3.5 44.0 ± 6.8 17:18c 20.0 ± 5.6 30.4 ± 5.0 25.4 ± 4.1 21.2 ± 5.9 22.7 ± 3.9 18.6 ± 2.3 20.0 ± 1.9 18.7 ± 3.3 16.4 ± 3.2 18.2 ± 3.7 18:36 GLA 1.1 ± 0.4 1.2 ± 0.2 0.8 ± 0.3 1.0 ± 0.1 0.7 ± 0.1 1.4 ± 0.2 0.4 ± 0.2 0.9 ± 0.3 0.6 ± 0.1 1.1 ± 0.0 18:43 3.4 ± 0.8 3.1 ± 0.7 3.5 ± 0.5 2.6 ± 0.6 3.3 ± 0.4 4.4 ± 1.2 2.9 ± 0.4 3.3 ± 0.6 2.4 ± 0.2 3.0 ± 0.5 18:2 1.6 ± 0.7 0.5 ± 0.3 4.0 ± 0.5 0.4 ± 0.2 3.4 ± 0.3 0.9 ± 0.9 2.7 ± 0.4 1.2 ± 1.1 1.5 ± 0.2 0.6 ± 0.4 18:26 99.8 ± 12.3 127.8 ± 19.1 115.1 ± 8.9 97.0 ± 19.4 82.4 ± 12.1 121.9 ± 11.4 87.1 ± 7.0 97.7 ± 11.4 73.6 ± 9.5 111.5 ± 13.6 18:33 ALA 15.0 ± 1.9 17.1 ± 2.0 20.8 ± 3.7 13.2 ± 3.5 18.8 ± 4.3 14.0 ± 1.8 17.6 ± 1.0 11.7 ± 2.3 16.3 ± 1.7 11.0 ± 1.8 18:19c OA 708.5 ± 161.6 579.8 ± 148.4 683.1 ± 147.0 464.3 ± 19.0 589.2 ± 183.0 523.0 ± 86.5 536.0 ± 83.3 561.6 ± 79.3 552.1 ± 101.2 684.5 ± 158.6 18:17c 44.3 ± 9.9 37.4 ± 6.6 94.5 ± 7.7 27.4 ± 7.5 65.3 ± 7.5 38.6 ± 10.5 63.6 ± 9.4 32.8 ± 10.3 38.9 ± 1.5 28.6 ± 5.4 18:17t 63.6 ± 17.5 56.3 ± 6.9 152.3 ± 16.7 35.7 ± 6.2 123.2 ± 15.4 42.6 ± 11.3 104.8 ± 19.0 43.2 ± 15.0 61.2 ± 7.5 40.6 ± 11.6 18:15c 6.1 ± 1.5 9.4 ± 0.7 10.6 ± 0.4 4.7 ± 1.2 8.4 ± 1.3 5.8 ± 1.0 8.8 ± 1.1 5.9 ± 1.6 7.9 ± 1.5 4.7 ± 1.2 18:0 370.9 ± 69.5 518.0 ± 83.8 464.3 ± 52.4 364.2 ± 109.3 365.3 ± 70.4 313.4 ± 41.3 376.8 ± 30.8 324.6 ± 54.4 344.8 ± 65.1 343.2 ± 81.9 CLA 8.6 ± 3.3 12.5 ± 1.9 10.7 ± 2.4 8.3 ± 3.7 8.5 ± 1.2 7.7 ± 2.4 9.4 ± 0.9 7.7 ± 2.2 9.2 ± 2.8 9.2 ± 1.9 20:46 ARA 32.7 ± 3.0 31.2 ± 6.7 25.8 ± 4.1 32.0 ± 6.8 16.1 ± 2.5 35.2 ± 2.8 19.0 ± 4.0 29.7 ± 2.8 20.4 ± 2.3 31.0 ± 2.9 20:53 EPA 10.0 ± 0.5 10.0 ± 3.1 11.4 ± 3.3 10.7 ± 2.2 7.5 ± 1.5 12.2 ± 1.8 8.9 ± 2.7 8.8 ± 1.4 7.5 ± 1.0 8.6 ± 2.1 20:36 4.2 ± 0.4 4.6 ± 0.9 4.0 ± 0.4 4.3 ± 0.7 2.6 ± 0.4 5.1 ± 0.5 3.5 ± 1.1 3.9 ± 0.3 2.8 ± 0.4 4.8 ± 0.4 20:1 1.3 ± 0.6 0.9 ± 0.4 2.2 ± 1.2 0.8 ± 0.4 1.5 ± 1.0 1.4 ± 0.5 2.5 ± 1.9 0.5 ± 0.3 0.7 ± 0.2 1.3 ± 0.2 20:19 3.5 ± 0.6 3.0 ± 0.5 5.9 ± 0.3 1.9 ± 0.6 4.5 ± 0.5 3.0 ± 1.1 4.6 ± 0.6 2.0 ± 0.9 2.8 ± 0.4 2.2 ± 0.6 20:17c 0.6 ± 0.3 0.5 ± 0.2 0.9 ± 0.3 0.0 ± 0.0 0.5 ± 0.1 0.3 ± 0.3 0.3 ± 0.3 0.3 ± 0.2 0.0 ± 0.0 0.2 ± 0.2 20:0 3.9 ± 0.9 4.6 ± 0.7 5.3 ± 0.3 3.2 ± 1.0 4.2 ± 0.7 3.2 ± 0.6 6.5 ± 0.8 3.5 ± 1.0 3.6 ± 0.5 2.8 ± 0.6 22:63 DHA 3.9 ± 0.5 3.3 ± 1.1 3.5 ± 0.8 3.4 ± 0.7 3.5 ± 1.1 6.9 ± 2.0 2.8 ± 0.7 3.1 ± 0.6 3.3 ± 0.2 3.9 ± 0.9 22:46 1.4 ± 0.2 1.3 ± 0.1 0.9 ± 0.3 1.3 ± 0.3 0.4 ± 0.2 1.5 ± 0.1 0.5 ± 0.3 1.3 ± 0.1 0.6 ± 0.3 1.6 ± 0.1 22:53 DPA 10.4 ± 0.7 9.5 ± 2.5 9.7 ± 2.2 10.0 ± 1.6 7.0 ± 1.3 10.8 ± 1.3 8.7 ± 1.7 9.1 ± 0.9 8.3 ± 0.8 9.0 ± 1.3 22:0 1.9 ± 0.2 1.8 ± 0.1 1.7 ± 0.1 1.8 ± 0.3 1.4 ± 0.4 2.1 ± 0.1 2.3 ± 0.6 1.5 ± 0.5 1.4 ± 0.1 1.8 ± 0.1 24:0 1.7 ± 0.1 1.7 ± 0.2 1.6 ± 0.2 1.4 ± 0.3 1.4 ± 0.4 1.3 ± 0.5 1.3 ± 0.2 1.3 ± 0.4 1.3 ± 0.1 1.7 ± 0.1 EPA+DHA 13.9 ± 0.9 13.3 ± 4.2 14.9 ± 4.0 14.1 ± 2.9 10.9 ± 2.6 19.1 ± 3.6 11.7 ± 3.3 11.9 ± 2.0 10.9 ± 1.2 12.5 ± 2.9 EPA+DPA+DHA 24.3 ± 1.6 22.8 ± 6.6 24.6 ± 6.2 24.1 ± 4.4 17.9 ± 3.8 29.9 ± 4.9 20.3 ± 4.9 21.0 ± 2.9 19.1 ± 1.9 21.5 ± 4.2 Total SFA 845.1 ± 171.4 1228.2 ± 190.9 1056.7 ± 125.5 870.0 ± 228.4 884.0 ± 160.0 799.0 ± 99.3 893.3 ± 61.3 761.0 ± 123.8 764.1 ± 119.7 797.1 ± 184.1 Total MUFA 895.4 ± 195.8 774.4 ± 150.7 1036.9 ± 171.5 596.8 ± 45.0 873.0 ± 220.4 677.7 ± 77.5 789.6 ± 108.7 700.0 ± 115.4 719.2 ± 113.4 813.6 ± 185.7 Total PUFA 208.0 ± 20.8 233.6 ± 24.0 231.9 ± 26.1 192.4 ± 37.9 174.8 ± 27.8 232.0 ± 24.8 182.9 ± 16.0 187.3 ± 23.2 161.1 ± 13.8 204.1 ± 21.4 Total Omega 3 42.9 ± 3.4 43.1 ± 7.7 49.1 ± 9.8 40.0 ± 7.6 40.8 ± 8.3 49.5 ± 7.9 41.3 ± 6.0 36.9 ± 5.2 37.8 ± 3.4 35.7 ± 5.7 Total Omega 6 139.1 ± 14.3 166.0 ± 20.7 146.5 ± 12.6 135.7 ± 25.8 102.0 ± 14.8 165.2 ± 14.5 110.6 ± 12.2 133.5 ± 14.5 98.0 ± 10.4 150.1 ± 13.6 6 : 3 Ratio 3.2 ± 0.1 4.2 ± 0.9 3.2 ± 0.4 3.4 ± 0.1 2.6 ± 0.3 3.5 ± 0.3 2.7 ± 0.2 3.7 ± 0.3 2.6 ± 0.1 4.3 ± 0.5 Total IMF % 3.1 ± 0.6 4.3 ± 0.6 3.9 ± 0.5 3.1 ± 0.3 4.3 ± 0.8 3.4 ± 0.8 3.8 ± 0.4 3.1 ± 0.2 3.5 ± 1.0 2.7 ± 0.6

Sire Breed and Supplement

Texel Canola (4) Lupin (4) Canola (4)

Coopworth

Lupin (4) Canola (4) Lupin (4)

White Suffolk Canola (3) Lupin (3) Dorset East Friesian

Canola (4) Lupin (4)

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Table 2.5 Mean intramuscular fatty acid content (±standard errors) for sex and supplement type in mg / 100g of the longissimus dorsi.